基于高通量优化的重组人血白蛋白纯化过程研究

项目名称： 基于高通量优化的重组人血白蛋白纯化过程研究

项目编号： No.21476198

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 有机化学

项目作者： 林东强

作者单位： 浙江大学

项目金额： 90万元

中文摘要： 针对临床药用重组人血白蛋白（rHSA）分离纯化的高要求，引入高通量的过程开发和优化方法，提高过程开发效率，实现分离过程的最优化。一方面，构建适用于层析分离优化的高通量过程开发方法，显著提高实验通量和优化空间，探讨酵母发酵液中复杂杂质的有效去除，实现单级层析优化和多级优化组合；另一方面，针对难分离的rHSA结合杂质，提出置换分离新概念，借助计算机分子模拟的高通量筛选，优化设计置换分子，组合疏水和静电作用，协调疏水结合和静电排斥解离，实现rHSA结合杂质的置换分离；在此基础上，优化rHSA分离流程，提高过程整体效率，实现高纯rHSA制备。本项目将建立基于高通量过程开发和分子模拟辅助设计的研究新方法，为生物分离过程优化和新方法构建提供指导，可推广到其它药用蛋白和生物活性物质的分离纯化过程。

中文关键词： 生物分离；高通量过程开发；重组人血白蛋白；层析；过程设计

英文摘要： Aiming at the requirement of high purity of recombinant human serum albumin (rHSA) for the pharmaceutical applications, new strategy based on the high-throughput process development (HTPD) and optimization would be introduced and used to improve the efficiency of process development. A HTPD-based platform would be established for the development and optimization of chromatographic separation of rHSA. The experimental factors and the screening space would be enhanced significantly. The optimized separation conditions for single-step chromatography and the combination of several chromatographic steps would be studied. The complicated impurities in the yeast culture broth would be removed efficiently. On the other hand, new concept of displacement separation would be proposed for the purification of the binding impurities on rHSA. With the computer-aided molecular simulation methods, the structure of displacement molecules would be designed and analyzed. The hydrophobic interactions would be forcued to improve the binding of displacement molecule onto rHSA and the electrostatic repulsion would be used for the departure of displacement molecule from rHSA. With the displacement molecules designed, the binding impurities on rHSA would be removed significantly. Based on the separation of the fermentation impurities and the binding impurities on rHSA, the downstream process would be designed for the production of rHSA with high purity from the yeast culture broth. The process efficiency would be improved significantly. In this project new methods based on HTPD and molecular simulation-aided design would be established, which would certainly improve the optimization of downstream process and develop new process for bioseparation.

英文关键词： Bioseparation;High-throughput process development;recombinant human serum albumin;chromatography;process design